Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos
Poria cocos (P. cocos) has long been used as traditional Chinese medicine and triterpenoids are the most important pharmacologically active constituents of this fungus. Farnesyl pyrophosphate synthase (FPS) is a key enzyme of triterpenoids biosynthesis. The gene encoding FPS was cloned from P. cocos...
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doaj-2ea969d5c38847b4b5f7e7af9da87f422020-11-24T22:59:56ZengMDPI AGInternational Journal of Molecular Sciences1422-00672014-12-011512221882220210.3390/ijms151222188ijms151222188Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocosJianrong Wang0Yangyuan Li1Danni Liu2Guangdong VTR Bio-Tech Co., Ltd., Zhuhai 519060, Guangdong, ChinaGuangdong VTR Bio-Tech Co., Ltd., Zhuhai 519060, Guangdong, ChinaGuangdong VTR Bio-Tech Co., Ltd., Zhuhai 519060, Guangdong, ChinaPoria cocos (P. cocos) has long been used as traditional Chinese medicine and triterpenoids are the most important pharmacologically active constituents of this fungus. Farnesyl pyrophosphate synthase (FPS) is a key enzyme of triterpenoids biosynthesis. The gene encoding FPS was cloned from P. cocos by degenerate PCR, inverse PCR and cassette PCR. The open reading frame of the gene is 1086 bp in length, corresponding to a predicted polypeptide of 361 amino acid residues with a molecular weight of 41.2 kDa. Comparison of the P. cocos FPS deduced amino acid sequence with other species showed the highest identity with Ganoderma lucidum (74%). The predicted P. cocos FPS shares at least four conserved regions involved in the enzymatic activity with the FPSs of varied species. The recombinant protein was expressed in Pichia pastoris and purified. Gas chromatography analysis showed that the recombinant FPS could catalyze the formation of farnesyl diphosphate (FPP) from geranyl diphosphate (GPP) and isopentenyl diphosphate (IPP). Furthermore, the expression profile of the FPS gene and content of total triterpenoids under different stages of development and methyl jasmonate treatments were determined. The results indicated that there is a positive correlation between the activity of FPS and the amount of total triterpenoids produced in P. cocos.http://www.mdpi.com/1422-0067/15/12/22188farnesyl diphosphate synthasemethyl jasmonate Poria cocostriterpenoids |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jianrong Wang Yangyuan Li Danni Liu |
spellingShingle |
Jianrong Wang Yangyuan Li Danni Liu Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos International Journal of Molecular Sciences farnesyl diphosphate synthase methyl jasmonate Poria cocos triterpenoids |
author_facet |
Jianrong Wang Yangyuan Li Danni Liu |
author_sort |
Jianrong Wang |
title |
Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos |
title_short |
Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos |
title_full |
Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos |
title_fullStr |
Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos |
title_full_unstemmed |
Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos |
title_sort |
cloning and characterization of farnesyl diphosphate synthase gene involved in triterpenoids biosynthesis from poria cocos |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2014-12-01 |
description |
Poria cocos (P. cocos) has long been used as traditional Chinese medicine and triterpenoids are the most important pharmacologically active constituents of this fungus. Farnesyl pyrophosphate synthase (FPS) is a key enzyme of triterpenoids biosynthesis. The gene encoding FPS was cloned from P. cocos by degenerate PCR, inverse PCR and cassette PCR. The open reading frame of the gene is 1086 bp in length, corresponding to a predicted polypeptide of 361 amino acid residues with a molecular weight of 41.2 kDa. Comparison of the P. cocos FPS deduced amino acid sequence with other species showed the highest identity with Ganoderma lucidum (74%). The predicted P. cocos FPS shares at least four conserved regions involved in the enzymatic activity with the FPSs of varied species. The recombinant protein was expressed in Pichia pastoris and purified. Gas chromatography analysis showed that the recombinant FPS could catalyze the formation of farnesyl diphosphate (FPP) from geranyl diphosphate (GPP) and isopentenyl diphosphate (IPP). Furthermore, the expression profile of the FPS gene and content of total triterpenoids under different stages of development and methyl jasmonate treatments were determined. The results indicated that there is a positive correlation between the activity of FPS and the amount of total triterpenoids produced in P. cocos. |
topic |
farnesyl diphosphate synthase methyl jasmonate Poria cocos triterpenoids |
url |
http://www.mdpi.com/1422-0067/15/12/22188 |
work_keys_str_mv |
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